论文部分内容阅读
目的 :构建mBMP 4真核表达载体并转染成纤维细胞 ,观测外源基因在靶细胞内的表达及对细胞表现型的影响。方法 :利用基因重组技术构建mBMP 4基因真核表达载体 pCR 3 .1uni mBMP 4;利用基因转染技术体外转染mBMP 4基因至成纤维细胞NIH 3T3 ,转染的细胞用G418筛选 ,利用斑点杂交和免疫组化检测转染 4周后外源基因表达的情况。结果 :琼脂糖电泳证明mBMP 4真核表达载体 pCR 3 .1uni mBMP 4构建成功。细胞转染后第 4周 ,斑点杂交和免疫组化均显示为阳性 ,阳性细胞碱性磷酸酶表达升高。结论 :外源BMP 4基因可在成纤维细胞内获得表达并可影响靶细胞的表现型。
OBJECTIVE: To construct mBMP 4 eukaryotic expression vector and transfect it into fibroblasts to observe the expression of exogenous gene in target cells and its effect on cell phenotype. METHODS: The mBMP 4 gene eukaryotic expression vector pCR 3 .1 uni mBMP 4 was constructed by gene recombination technique. The mBMP 4 gene was transfected into fibroblasts NIH 3T3 by gene transfection. The transfected cells were selected by G418. Immunohistochemistry was used to detect the expression of foreign genes 4 weeks after transfection. Results: The agarose gel electrophoresis proved that mBMP 4 eukaryotic expression vector pCR 3 .1uni mBMP 4 was successfully constructed. At 4 weeks after transfection, dot blot and immunohistochemistry showed positive staining and positive cells showed elevated alkaline phosphatase expression. Conclusion: Exogenous BMP 4 gene can be expressed in fibroblasts and can affect the target cell phenotype.